Identification Of Interaction And Expression Level Of MADS In Hickory (Carya Cathayensis Sarg.)

Flowering event is a transitive stage in plant from vegetative phase to the reproductive phase whichis controlled by diverse endogenous and environmental factors. The floral MADS-box genes playcrucial roles in the whole stages of plant development. Many studys demonstrate that MADStranscription factors function in a combinational manner. In this study, we obtained several ORFs ofMADS based on the454transcroptome sequencing of pistilate in Carya cathayensis Sarg. Thepreliminary research results are as following:1. According to alignment and phylogenetic analyses, it demonstrated that they were allMIKC-type MADS-box genes of plants, named as CcAP1(AHI85952), CcSVP (AHI85951), CcSOC1(AHI85950), CcTM，respectively. And all these MADS-box transcription factors shared a highlyconserved MADS-domain (about59aa) and a comparable conserved K-domain.2. qRT-PCR of CcTM in different tissues indicated that CcTM could play role in flowering inhickory, which expression level in floral buds7-9times to in leaves and vegetative buds.3.H2Y assays showed that the combinations of CcAP1/CcAP1, CcAP1/CcSVP, CcAP1/CcFLC1,CcAP1/CcFLC2, CcSVP/CcSVP, CcSVP/CcFLC1, CcFLC1/CcFLC1, CcFLC1/CcFLC2,CcFLC2/CcFLC2had interaction with each other. However, CcSOC1had performed considerable “interia” inour experiment.4. K-domain played a pivotal role in dimerization of CcSOC1. CcSOC1protein might potentiallyform homo/hetero-dimer with K-domain, though M-domain appeared to be a hinder for CcSOC1dimerization according to our assays.5. Western blot of mated yeast conformed that target proteins did express in both unmated yeastand mated yeast, moreover, the level of protein expression was sufficient for positive phenomenon if theprotein had interaction with others.6. Western blotting showed that the expression level of CcSVP in different time in male or femalebuds presented a “peak” appearance. Native page and blotted against the antibody of CcSVPdemonstracted that CcSVP forms higher-order complexes, either homo or/and hetero-complexes inhickory flowering buds.